【作者】 马琼；

【导师】 黄建国；

【作者基本信息】 西南农业大学 ， 微生物学， 2004， 硕士

【摘要】 马尾松生长快，适应性强，用途广，经济价值高，是我国尤其是西南地区人工林和天然林的主要树种。但是，在植树造林工作中，马尾松苗木成活率低，生长缓慢，土传病害严重。因此，培育高质量的林木种苗成为保证造林质量的重要环节。 大量的研究表明，利用优良的外生菌根真菌进行苗床接种，培育菌根化林木种苗可以增加苗木吸收养分和水分，提高它们的抗逆性(抗旱、抗病)，促进生长发育。目前，苗床接种优良的外生菌根真菌，培育菌根化苗木已经在国外广泛应用于林木种苗生产，我国也有少量应用。必须指出，生产菌根化苗木的关键是选用适宜当地生态条件的优良菌株，用于接种，培育出菌根化林木种苗。 试验从马尾松大面积分布的三峡库区，分离出代表当地生态条件的松乳菇(Lactarius deliciosus 1、2和3)，并选择中国南方具有代表性的华南牛肝菌(Suillus placidu 504)和双色蜡蘑(Laccaria bicolor Lb S238N)，研究了松乳菇3个菌株的培养特性，并利用Lb S238N、Sp 504、Ld01和Ld03苗床接种马尾松，观察了它们对幼苗的生长和养分吸收的影响，目的是为三峡库区马尾松的植树造林提供培育菌根化苗木技术。试验结果如下： 1．用Pachlewski和PDA培养基分离Lactarius deliciosus，PDA的培养生长状况优于Pachlewski。在PDA培养基上，Lactarius deliciosus子实体不同部位的组织均能萌发长出正常菌丝，说明不同部位的组织都可用于分离培养。根据菌丝分泌物的颜色，Lactarius deliciosus可以分为3个菌株，分别命名为Ld 01、Ld 02和Ld 03。 2．在分离菌株的扩大培养过程中，PDA培养基的培养效果仍然较好，不仅菌丝的生长速度快，而且长势好；但在Pachlewski培养基上的菌丝生长速度缓慢，长势差。说明PDA培养基用于分离和培养Lactarius deliciosus的效果优于Pachlewski培养基。比较三株Lactarius deliciosus菌株在PDA培养基上的生长情况，Ld 01长势最强，Ld 02和Ld 03差异不大。 3．在制备生产用菌种时，较佳的培养基组成和培养条件是：温度25-28℃，pH5.0-6.0，固体基质(珍珠岩和蛭石，V／V=1：1)，用PDA培养液拌料，含水量65-70％。用三角瓶、食用菌栽培袋和罐头瓶三种容器制生产用菌剂，菌丝在三角瓶中生长西南农业大学硕士学位论文摘要最快，在接种袋和罐头瓶中生长较慢。一般而言，菌丝长满固体培养基需要25-40d，Lb 5238N、Ldol和Ld03菌丝生长速度较快，SP 504生长速度较慢，Ld02最差。4.接种外生菌根真菌，可以观察到幼苗根系结构改变，根毛消失，在根尖表面布满菌丝，菌根感染率超过65%以上，较对照均达显著水平(P<0 .05)。其中，以Lb5328N和SP 504的感染率最高，达到75%以上。此外，接种外生菌根真菌之后，苗木高度、地径和生物量等显著增加。就促进幼苗生长而言，Lb s328N和SP 504的效果较好，优于在当地分离的三株Lactarius deliciosus。Lb 5328N和SP 504对苗木各生长指标的影响均达显著水平(P<0.05)。幼苗株高、地径和生物量分别比对照(不接种)增加了72.41%、34.56%和124.56%(L b 5328N);82.65%、22.57%和107.02%(SP 504)O5.苗床接种外生菌根真菌，幼苗的养分含量显著提高。与对照(不接种)相比，接种菌根真菌后，幼苗地上部和地下部的含氮量平均提高了17.83%一31 .80%和11.17%一26.08%，差异达显著水平(P<0.05);含磷量提高了32.11%一44.46%和2623%一0.79%，差异达极显著水平(P<0 .01);含钾量提高了15.81%一38.66%和12.62%一23.33%。 综上所述，苗床接种外生菌根真菌之后，养分吸收量增加，促进幼苗生长，说明培育菌根化化苗木是提高马尾松种苗质量的实用技术。更多还原

【Abstract】 Pinus Massoniana, the main tree species in natural and artificial forests in China, especially in Southwest China, grows fast and adapts to variable conditions. The tree is used economically for multi-purposes. Pinus Massoniana seedlings are difficult to be survival, slow in growing and serious in soil born disease. The quality of the seedlings is necessary to be improved in artificial forestation.It was reported that the absorption of nutrients and water by seedlings could be promoted by inoculation of ectomycorrhizal fungi. The abilities of stress resistance, including drought and disease, could also be increased by this way, resulting the fast growing of seedlings. Inoculation of ectomycorrhizal fungi in nurse bed to produce ectomycorrhizal seedlings has been widely adopted abroad, but little in China, if any. It is necessary to point out that the inocunation of the ectomycorrhizal fungi suitable to local natural conditions is important in producing ectomycorhizal seedlings.In this experiment, typical ectomycorhizal fungi strains(Lactarius delicious 01, 02 and 03) were isolated from Three-Gorges Reservoir where Pinus Massoniana was very popular. And Suillus placidu 504(sp 504) and Laccaria bicolor S238N (Lb 23 8N) typical in South China were also chosen to study the effects of ectomycorrhizal fungi on the growth and nutrient absorption of Pinus Massoniana. And the culture characteristics of three strains of Lactarius delicious were also studied. The object of this study is to supply techniques in culturing ectomycorrhizal seedlings for planting trees in Three-Gorges Reservoir. The results are as follows:1. Growth of Lactarius deliciosus on PDA was better than that on Pachlewski. All selected tissues from different positions of the same Lactarius deliciosus basidiocarp can germinate mycelium on PDA. It showed that different position tissues could be used in isolation culture. According to the secretion of mycelium, Lactarius deliciosus strains were classified into Ld 01,Ld 02 and Ld 032. The isolated strains had better growth potential on PDA, and mycelium grew quickly and strongly. While they grew slowly and poorly on Pachlewski. It indicated that PDA was better in isolation and culturing Lactarius deliciosus than Pachlewski. The growth potentials of the three isolated strains were very different on PDA,Ld 01 was best,and no significant difference was found between Ld 02 and Ld 03.3. Better culture material components and conditions in preparing for inocula were as follows: 25-28℃ of temperature, PH value with 5.0-6.0, usage of PDA as culturing material, 65-70% of water content, solid material of perlite and vermiculite (V/V=1:1). The mycelium grew fast in flask and slowly in edible fungi planting sack and can. Generally, the time of solid material overgrowing with mycelium was about 25-40d, and the growth of mycelium of Lb S238N, Ld 01 and Ld 03 was fast, SP 504 was slow and Ld 02 was the worst.4.After inoculation with ectomycorrhizal, the root structure of infected seedlings was deformed, and no root hair was observed. Mycelium overgrew root tip. The infection rate was more than 65%, of which Lb S328N and Sp 504 were the highest, reaching 75%. Moreover, growth height, basal diameter and biomass of inoculated seedlings were also increased significantly. Lb S328N and Sp 504 increased the growth of seedlings more than three isolated Lactarius deliciosus strains. Compared with the control, the increments of height, basal diameter and biomass of seedlings of Lb S328N were 72.41%, 34.56% and 124.56%, respectively, and that of Sp 504 were 82.65%, 22.57% and 107.02%, respectively.5.The concentrations of N, P and K were significantly increased after inoculation. Compared with the control, the increments of N in underground and aboveground parts in inoculated seedlings were 17.83%-31.80% and 11.17%-26.08%( P<0.05), those of P were 32.11%-44.46% and 26.23%-40.79%( P<0.01), and those of K were 15.81%-38.66% and 12.62%-23.33%.In conclusion, ectomycorrhizal fungus inoculation can increase nut更多还原